Thesis Working Plan
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Distribution, Genetic Characterization, and Life History of the James spinymussel, Pleurobema collina (Bivalvia: Unionidae), in Virginia and North Carolina by Melissa A. Petty Thesis submitted to the Faculty of the Virginia Polytechnic Institute and State University in partial fulfillment of the requirements for the degree of Master of Science in Fisheries and Wildlife Sciences Approved: ____________________________ _____________________________ Dr. Richard J. Neves, Chairperson Dr. Eric M. Hallerman _________________________ Dr. Paul Angermeier February 11, 2005 Blacksburg, Virginia Keywords: James spinymussel, Pleurobema collina, Distribution, Survey, Life History, DNA Sequences, Fish Host, Phenotype, Morphology, Monophyletic, Genetic Drift Distribution, Genetic Characterization, and Life History of the James spinymussel, Pleurobema collina (Bivalvia: Unionidae), in Virginia and North Carolina by Melissa A. Petty Richard J. Neves, Chairperson Fisheries and Wildlife Sciences (ABSTRACT) Three spined, mussel species occur in the United States along the Atlantic slope; James spinymussel (Pleurobema collina), Tar spinymussel (Elliptio steinstansana), and Altamaha spinymussel (E. spinosa). The James spinymussel was listed as endangered in 1988, and was until recently considered to be endemic to the James River basin (Clarke and Neves 1984; USFWS 1990). Biologists with the North Carolina Department of Transportation (NCDOT) discovered spinymussel populations in the Dan and Mayo rivers in NC in 2000 and 2001, respectively. The U.S. Fish & Wildlife Service (USFWS) tentatively identified this species as Pleurobema collina. My project proposed by the Virginia Cooperative Fish and Wildlife Research Unit to the USFWS and the Virginia Transportation Research Council, determined where P. collina resides in VA and what the extent of its range is within the state. An informal preliminary survey design for P. collina was used during the summer of 2002 and simple random sampling was deployed in 2003-2004 surveys to provide a good basis for comparison to gauge the efficiency of the informal sampling design. In 2002, a total of 116 person-hours were spent surveying 39 localities on the Mayo, Dan, and Smith rivers. A total of 96 P. collina was observed in the South Fork of the Mayo River, Patrick and Henry counties, VA. A documented range of 24 rkm was established in the South Fork Mayo River. During the summers of 2003 and 2004, a total of 228 person-hours were spent surveying 38 equal-area river reaches (10, 000 m2) on the mainstems of the Dan, Smith, South Mayo, and Banister rivers. No specimens of P. collina (live or relic shells) were detected. A simple random sampling approach was designed to be easy, relatively quick and cost effective, applicable to most rivers, and to provide actual numbers for comparison. Negative results were only reported after 6 person-hours of searching within each randomly selected, equal-area river reach had been expended. P. collina was declared absent from the VA random sites surveyed in 2003- 2004 with a confidence of ~90%. A genetic characterization of four extant populations of P. collina was conducted to assess its taxonomic affinity and to resolve conservation issues related to recovery planning and management actions. The populations were examined for phenotypic variation, and were characterized phylogenetically using DNA sequences. A comprehensive analysis was performed for both separate and combined mitochondrial (357 bp of cytochrome-b, 916 bp of ND-1) and nuclear (502 bp of ITS-1) DNA sequences. Based on comprehensive molecular, morphological, and life history data, populations of P. collina sampled from the Dan River sub-drainage do not warrant separate species designation from P. collina sampled from the James River drainage. ii Acknowledgments I would first like to thank my advisor, Richard J. Neves, for his support and guidance over the past three years. You have given me the opportunity to study endangered unionids and also introduced me to an area of science conservation that I hope will be a part of the rest of my life. You have given me the resources and freedom to fully pursue a graduate education dedicated to the conservation of imperiled species. I also would like to thank my other committee members, Eric M. Hallerman and Paul L. Angermeier, for their advice and patience. Special thanks to Eric Hallerman for providing the education and guidance I craved in the genetics work of this project, and for making his laboratory available to me. Your skill, passion, and compassion as a teacher are unsurpassed. I also would like to extend a special thanks to Paul Angermeier for his guidance in transforming my survey work into real science. Paul provided many valuable comments on the thesis manuscript. I am very grateful for all of the immeasurable advice and guidance and the ways they have improved my life as a scientist. I would like to thank biologists Tim Savidge and Sharon Snyder of The Catena Group for their friendship and help on the fieldwork for this project. Many conversations with you guys on the banks of the Dan River after long days in the field served as the catalyst to investigate the taxonomy of the James spinymussel. I would like to thank my colleague Jess Jones for his encouragement and patience in the genetics laboratory and also for photographing the James spinymussel in the wild. I would especially like to thank Nathan Johnson, Dan Dutton, David Garst, Rachel Mair, and Jeremiah Purdum for your skill and dedicated hard work in the field and laboratory. Without your help, this project could not have been completed. A special thank you goes to Jennifer Struthers, iii Nathan Johnson, and Kathy Finne, for teaching me the methods and techniques of the genetics laboratory. I would like to thank Jeanne Serb (University of California) for kindly sharing DNA sequences and PCR recipes for the ND1 region of the mitochondrial genome. Additional thanks goes to Art Bogan (University of Alabama) for providing tissue samples of Elliptio steinstansana and Pleurobema collina. Financial support for this project was provided by the Virginia Transportation Research Council (VTRC) and the U.S. Fish and Wildlife Service (USFWS). I would like to thank the now retired John Alderman (North Carolina Wildlife Resources Commission) for supporting freshwater mollusk recovery and conservation throughout most of his career. I would especially like to thank Laura Roghair, Conservation Management Institute (CMI) and Brian Watson, Virginia Department of Game and Inland Fisheries (VDGIF) for creating and providing each and every map for this project. I would like to thank the many biologists who helped on various aspects of this project including Mike Pinder (VDGIF), John Schmerfeld, and John Fridell (USFWS). A special thanks goes to Penelope Pooler for her invaluable knowledge and skill with the statistics of this project. The support and aid of my fellow graduate students was sincerely appreciated. Your interest helped to develop my project for the better. I also thank the private landowners of the Dan River and Banister River drainages. Without your cooperation and interest, much of my work could not have been completed. I want to thank my parents, Dorothy and David Petty, for making my time at Virginia Tech possible. Your love, support, and guidance through all of my academic efforts will never be forgotten and always appreciated. Finally, I would like to thank John, Cora, Andrew, Lora, Colleen, Mary, Kate, and Sarah; only through your love, good food, and good wine could I have made it to the end of my Master of Science degree. iv Table of Contents Chapter 1. Range of Pleurobema collina in the Roanoke River System……………...1 Introduction………………………………………………………………………………..4 Newly-discovered distribution in the Dan River sub-basin…….…………………4 Survey in the James River Drainage, VA…………………………………………5 Survey and current distribution in the Dan River sub-basin, NC…………………6 Informal vs. probability-based survey design……………………………………..9 Study area …………………………………………………………………………….…12 Roanoke River Basin..…………………………………………………………...12 Dan River sub-basin in the Roanoke River ……………………………………..14 Methods…………………………………………………………………………………15 Informal preliminary survey in Roanoke River Drainage, VA………………….15 Simple random survey in the Roanoke River Drainage, VA……………………16 Results …………………………………………………………………………………..21 Informal preliminary survey in the Roanoke River Drainage, VA ……………..21 Simple random survey in the Roanoke River Drainage, VA……………………22 Discussion ………………………………………………………………………………24 Conclusions and management implications……………………………………..27 Literature Cited………………………………………………………………………….31 Chapter 2. Genetic Characterization and Life History of the Endangered James Spinymussel Pleurobema collina (Bivalvia: Unionidae)…………………………….64 Introduction …………………………………………………………………………….67 Historical classification and characteristics of the James spinymussel…………70 v Identification of freshwater mussel conservation units………………………….72 Designation of conservation units……………………………………………….75 Methods………………………………………………………………………………….76 Shell material…………………………………………………………………….76 Tissue collection…………………………………………………………………76 DNA sequences …………………………………………………………………77 Length of glochidia and fecundity……………………………………………….78 Anatomy (mantle and papillae)…………………………………………………..79 Fish host specificity……………………………………………………………...79 Data analysis……………………………………………………………………..81 Results……………………………………………………………………………………83 Valve characters………………………………………………………………….83